An indirect immunofluorescence assay using a cell culture-derived antigen for detection of antibodies to the agent of human granulocytic ehrlichiosis.

An indirect immunofluorescence assay for the detection of human antibodies to the agent of human granulocytic ehrlichiosis (HGE) was developed and standardized. Antigen was prepared from a human promyelocytic leukemia cell line (HL-60) infected with a tick-derived isolate of the HGE agent (USG3). Suitable antigen presentation and preservation of cellular morphology were obtained when infected cells were applied and cultured on the slide, excess medium was removed, and cells were fixed with acetone. Use of a buffer containing bovine serum albumin and goat serum reduced background fluorescence, and use of an immunoglobulin G (gamma-specific) conjugate reduced nonspecific binding. The assay readily detected specific antibody from HGE patients and did not detect antibody from healthy individuals. No significant reactivity was noted in sera from patients with high titers of antibodies to other rickettsial species. We were able to identify antibodies reactive to USG3 antigen in samples from areas where HGE is endemic that had tested negative to other rickettsial agents. Animal sera reactive against Ehrlichia equi or Ehrlichia phagocytophila bound to the HGE antigen, indicating that the assay may be useful for veterinary use. Comparability between two different laboratories was assessed by using coded human sera exchanged between laboratories. Results from the two laboratories were similar, indicating that the assay can be easily integrated into use for routine testing for HGE. The assay was then compared to an assay using horse neutrophils infected with ehrlichiae. The two assays gave comparable results, indicating that the cell culture-derived antigen can be used for testing samples that have been previously tested with E. equi as an antigen. The new assay offers several advantages over other immunofluorescence methods that use animal-derived antigen and is suitable for use in testing for human antibodies to the HGE agent.     

Bipolar recording of short-latency somatosensory evoked potentials after median nerve stimulation

Generators of median short-latency somatosensory evoked potentials were studied with three orthodiagonal pairs of bipolar electrodes. N11 was attributed to the dorsal root and dorsal column volleys. N13 had at least two subcomponents, generator dipoles of which are directed horizontally (N13a) and axially (N13b). N13a was generated in the lower cervical cord. N13b (bipolar) and P14 far-field (noncephalic reference) appeared to originate in the cuneate nucleus or spinocerebellar tracts as well as in the medial lemniscus. Bipolar recordings were useful in localizing cervical cord lesions, which was impossible in conventional monopolar recordings.     

Ipsilateral blepharospasm and contralateral hemidystonia and parkinsonism in a patient with a unilateral rostral brainstem-thalamic lesion: structural and functional abnormalities studied with CT, MRI, and PET scanning

A patient developed progressive right hemidystonia in childhood. Subsequently, left-sided blepharospasm, slurred and stuttering speech, and right-sided rigidity and bradykinesia, responsive to dopamine agonists, appeared. Investigation with computed tomography and magnetic resonance imaging (MRI) at age 43 years revealed a left-sided calcified rostral brainstem-thalamic lesion of uncertain aetiology. Although no structural lesion was seen in the striatal regions, L-[18F]-fluorodopa uptake was severely diminished in the left striatum but normal on the right. Dopamine receptor binding identified by [11C]-methylspiperone was in the normal range on both sides.     

Noninvasive assessment of upper extremity and hand ischemia

The diagnosis and follow-up of patients with ischemic problems of the arm and hand is facilitated by the selective use of simple noninvasive methods readily available in most vascular laboratories. These tests help differentiate between obstruction and vasospasm and document the severity of the circulatory impairment. Although they may not establish etiology, they often clarify the need for further laboratory tests or arteriography.     

FM sonography in diffuse liver disease: prospective assessment and blinded analysis

FM sonography - a signal-processing technique that uses frequency and phase information as well as amplitude data - shows promise in evaluation of patients with diffuse liver disease. In a prospective blinded review of 37 patients with biopsy-proved liver disease and 42 healthy volunteers, FM sonography was clearly superior to traditional amplitude-based (AM) sonography in distinguishing healthy from diseased subjects. Statistically significant differences were seen in accuracy (FM, 98.7%; AM, 84.8%), sensitivity (FM, 97.3%; AM, 70.3%), and negative predictive value (FM, 97.7%; AM, 78.8%). Our data also suggest that current FM sonographic techniques cannot differentiate among histologic findings associated with different hepatic parenchymal abnormalities. It is unclear, therefore, whether FM imaging can reduce the numbers of patients who require biopsy for diagnosis or the frequency of biopsy procedures in patients with known disease.     

Anaphylatoxin levels in human aqueous humor

Radioimmunoassay was used to measure levels of C3a, C4a, and C5a in aqueous humor from 13 normal eyes, 8 noninflamed eyes with a history of surgery or inflammation, and 14 eyes with anterior uveitis. The authors were unable to measure levels of C3a, C4a, or C5a in normal aqueous humor. In noninflamed aqueous humor from eyes with a history of surgery or inflammation, the authors were unable to measure levels of C4a or C5a, but were able to measure low levels of C3a in 3/8 patients. In aqueous humor from eyes with anterior uveitis, the authors were able to measure levels of C3a in all 14 patients, C4a in 9/14 patients, and C5a in 5/14 patients. Patients with severe anterior uveitis had higher levels of C3a than those with moderate anterior uveitis. The higher ratios of anaphylatoxin to protein levels in inflamed aqueous humor, when compared to normal plasma or noninflamed aqueous humor, suggested that complement was being activated by either the classical or alternative pathways in inflamed aqueous humor. Measurable levels of C3a without detectable C4a in five patients with anterior uveitis suggested alternative pathway activation of complement.     

Familial and sporadic hyperinsulinism: histopathologic findings and segregation analysis support a single autosomal recessive disorder

We evaluated the possible genetic contribution to hyperinsulinism in a series of patients seen during the past 15 years. Of 26 families, 5 (19%) had more than one child affected (multiplex family). There were no apparent differences between patients in the 5 multiplex and 21 simplex families, clinically, biochemically, or on histologic examination of the pancreatic specimens. The families studied had a total of 63 offspring; the 26 index patients had 37 siblings, 6 of whom were affected. After four patients with hyperinsulinism caused by adenoma were excluded from the study, segregation analysis was carried out to test the data for agreement with results expected if familial and isolated hyperinsulinism represented a single disease with recessive mode of inheritance and a segregation ratio of 0.25. Excellent agreement was found between the observed number of affected siblings (20) and the expected number (19.65), with a segregation ratio of 0.254. The results were consistent with the hypothesis that in most or all cases, hyperinsulinism is inherited as an autosomal recessive disease. There was no evidence of distinct familial and sporadic types.